Targeting Mcl-1 for Radiosensitization of Pancreatic Cancers

AbstractIn order to identify targets whose inhibition may enhance the efficacy of chemoradiation in pancreatic cancer, we previously conducted an RNAi library screen of 8,800 genes. We identified Mcl-1 (myeloid cell leukemia-1), an anti-apoptotic member of the Bcl-2 family, as a target for sensitizing pancreatic cancer cells to chemoradiation. In the present study we investigated Mcl-1 inhibition by either genetic or pharmacological approaches as a radiosensitizing strategy in pancreatic cancer cells. Mcl-1 depletion by siRNA produced significant radiosensitization in BxPC-3 and Panc-1 cells in association with Caspase-3 activation and PARP cleavage, but only minimal radiosensitization in MiaPaCa-2 cells. We next tested the ability of the recently identified, selective, small molecule inhibitor of Mcl-1, UMI77, to radiosensitize in pancreatic cancer cells. UMI77 caused dissociation of Mcl-1 from the pro-apoptotic protein Bak and produced significant radiosensitization in BxPC-3 and Panc-1 cells, but minimal radiosensitization in MiaPaCa-2 cells. Radiosensitization by UMI77 was associated with Caspase-3 activation and PARP cleavage. Importantly, UMI77 did not radiosensitize normal small intestinal cells. In contrast, ABT-737, an established inhibitor of Bcl-2, Bcl-XL, and Bcl-w, failed to radiosensitize pancreatic cancer cells suggesting the unique importance of Mcl-1 relative to other Bcl-2 family members to radiation survival in pancreatic cancer cells. Taken together, these results validate Mcl-1 as a target for radiosensitization of pancreatic cancer cells and demonstrate the ability of small molecules which bind the canonical BH3 groove of Mcl-1, causing displacement of Mcl-1 from Bak, to selectively radiosensitize pancreatic cancer cells

Hybrid Quinoline-Thiosemicarbazone Therapeutics as a New Treatment Opportunity for Alzheimer’s Disease‒Synthesis, In Vitro Cholinesterase Inhibitory Potential and Computational Modeling Analysis

From MDPI via Jisc Publications RouterHistory: accepted 2021-10-27, pub-electronic 2021-10-30Publication status: PublishedAlzheimer’s disease (AD) is a progressive neurodegenerative disorder and the leading cause of dementia worldwide. The limited pharmacological approaches based on cholinesterase inhibitors only provide symptomatic relief to AD patients. Moreover, the adverse side effects such as nausea, vomiting, loss of appetite, muscle cramps, and headaches associated with these drugs and numerous clinical trial failures present substantial limitations on the use of medications and call for a detailed insight of disease heterogeneity and development of preventive and multifactorial therapeutic strategies on urgent basis. In this context, we herein report a series of quinoline-thiosemicarbazone hybrid therapeutics as selective and potent inhibitors of cholinesterases. A facile multistep synthetic approach was utilized to generate target structures bearing multiple sites for chemical modifications and establishing drug-receptor interactions. The structures of all the synthesized compounds were fully established using readily available spectroscopic techniques (FTIR, 1H- and 13C-NMR). In vitro inhibitory results revealed compound 5b as a promising and lead inhibitor with an IC50 value of 0.12 ± 0.02 μM, a 5-fold higher potency than standard drug (galantamine; IC50 = 0.62 ± 0.01 μM). The synergistic effect of electron-rich (methoxy) group and ethylmorpholine moiety in quinoline-thiosemicarbazone conjugates contributes significantly in improving the inhibition level. Molecular docking analysis revealed various vital interactions of potent compounds with amino acid residues and reinforced the in vitro results. Kinetics experiments revealed the competitive mode of inhibition while ADME properties favored the translation of identified inhibitors into safe and promising drug candidates for pre-clinical testing. Collectively, inhibitory activity data and results from key physicochemical properties merit further research to ensure the design and development of safe and high-quality drug candidates for Alzheimer’s disease

Monitoring of Cadmium and Lead Accumulation in Soil-Forage-Animal Continuum in Pasture Land Irrigated with Ground Water in Bhalwal, Punjab, Pakistan

Recent study was directed to check the accumulation of Cd and Pb in pasture land treated with ground water. In particular the transfer of cadmium and lead from soil to forages and in turn to animal (buffaloes) was conducted in Bhalwal, Punjab, Pakistan which comes under sub-tropical environmental conditions. The Cd and Pb concentration in selected samples was explored by atomic absorption spectrophotometer (AA-6300 Shimadzu Japan). The results depicted the concentration of cadmium in water, soil, forages, milk and hair of buffaloes was in the range of 0. 00320 – 0.00866 mgL-1, 1.9500 to 5.3000 mg/kg, 0.300 to 0.7100 mgkg-1, 0.1033 to 0.4133 mgL-1 and 0.037 to 0.0656 mg/kg, respectively. The lead concentration was ranged from 0.004 mg/L to 1.963 mgL-1 for water, 5.960 -13.600 mg/kg for soil, 0.293 to 2.570 mg/kg for forages, 0.2166 to 6.100 mg/L for milk and 0.0206 to 0.074 mg/kg for hair samples. Various indices (BCF, PLI, EF, DIM and HRI) were examined and results presented that PLI and EF of Cd, EF and HRI for Pb was above 1 indicating that metal was causing pollution while value of BCF and DIM was below one. If exposed for an extended period of time through feed, forages with a higher Cd and Pb content may harm animal\u27s cells, create respiratory issues and have an adverse effect on the animal\u27s kidney, liver and lungs

MGP Panel is a comprehensive targeted genomics panel for molecular profiling of multiple myeloma patients

PURPOSE: We designed a comprehensive multiple myeloma (MM) targeted sequencing panel to identify common genomic abnormalities in a single assay and validated it against known standards. EXPERIMENTAL DESIGN: The panel comprised 228 genes/exons for mutations, 6 regions for translocations, and 56 regions for copy number abnormalities (CNAs). Toward panel validation, targeted sequencing was conducted on 233 patient samples and further validated using clinical fluorescence in situ hybridization (FISH) (translocations), multiplex ligation probe analysis (MLPA) (CNAs), whole genome sequencing (WGS) (CNAs, mutations, translocations) or droplet digital PCR (ddPCR) of known standards (mutations). RESULTS: Canonical IgH translocations were detected in 43.2% of patients by sequencing, and aligned with FISH except for one patient. CNAs determined by sequencing and MLPA for 22 regions were comparable in 103 samples and concordance between platforms was R2=0.969. VAFs for 74 mutations were compared between sequencing and ddPCR with concordance of R2=0.9849. CONCLUSIONS: In summary, we have developed a targeted sequencing panel that is as robust or superior to FISH and WGS. This molecular panel is cost effective, comprehensive, clinically actionable and can be routinely deployed to assist risk stratification at diagnosis or post-treatment to guide sequencing of therapies

Effect of early tranexamic acid administration on mortality, hysterectomy, and other morbidities in women with post-partum haemorrhage (WOMAN): an international, randomised, double-blind, placebo-controlled trial

Background Post-partum haemorrhage is the leading cause of maternal death worldwide. Early administration of tranexamic acid reduces deaths due to bleeding in trauma patients. We aimed to assess the effects of early administration of tranexamic acid on death, hysterectomy, and other relevant outcomes in women with post-partum haemorrhage. Methods In this randomised, double-blind, placebo-controlled trial, we recruited women aged 16 years and older with a clinical diagnosis of post-partum haemorrhage after a vaginal birth or caesarean section from 193 hospitals in 21 countries. We randomly assigned women to receive either 1 g intravenous tranexamic acid or matching placebo in addition to usual care. If bleeding continued after 30 min, or stopped and restarted within 24 h of the first dose, a second dose of 1 g of tranexamic acid or placebo could be given. Patients were assigned by selection of a numbered treatment pack from a box containing eight numbered packs that were identical apart from the pack number. Participants, care givers, and those assessing outcomes were masked to allocation. We originally planned to enrol 15 000 women with a composite primary endpoint of death from all-causes or hysterectomy within 42 days of giving birth. However, during the trial it became apparent that the decision to conduct a hysterectomy was often made at the same time as randomisation. Although tranexamic acid could influence the risk of death in these cases, it could not affect the risk of hysterectomy. We therefore increased the sample size from 15 000 to 20 000 women in order to estimate the effect of tranexamic acid on the risk of death from post-partum haemorrhage. All analyses were done on an intention-to-treat basis. This trial is registered with ISRCTN76912190 (Dec 8, 2008); ClinicalTrials.gov, number NCT00872469; and PACTR201007000192283. Findings Between March, 2010, and April, 2016, 20 060 women were enrolled and randomly assigned to receive tranexamic acid (n=10 051) or placebo (n=10 009), of whom 10 036 and 9985, respectively, were included in the analysis. Death due to bleeding was significantly reduced in women given tranexamic acid (155 [1·5%] of 10 036 patients vs 191 [1·9%] of 9985 in the placebo group, risk ratio [RR] 0·81, 95% CI 0·65–1·00; p=0·045), especially in women given treatment within 3 h of giving birth (89 [1·2%] in the tranexamic acid group vs 127 [1·7%] in the placebo group, RR 0·69, 95% CI 0·52–0·91; p=0·008). All other causes of death did not differ significantly by group. Hysterectomy was not reduced with tranexamic acid (358 [3·6%] patients in the tranexamic acid group vs 351 [3·5%] in the placebo group, RR 1·02, 95% CI 0·88–1·07; p=0·84). The composite primary endpoint of death from all causes or hysterectomy was not reduced with tranexamic acid (534 [5·3%] deaths or hysterectomies in the tranexamic acid group vs 546 [5·5%] in the placebo group, RR 0·97, 95% CI 0·87-1·09; p=0·65). Adverse events (including thromboembolic events) did not differ significantly in the tranexamic acid versus placebo group. Interpretation Tranexamic acid reduces death due to bleeding in women with post-partum haemorrhage with no adverse effects. When used as a treatment for postpartum haemorrhage, tranexamic acid should be given as soon as possible after bleeding onset. Funding London School of Hygiene & Tropical Medicine, Pfizer, UK Department of Health, Wellcome Trust, and Bill & Melinda Gates Foundation

Implementation and testing of a parallel layer peeling algorithm

Due to the character of the original source materials and the nature of batch digitization, quality control issues may be present in this document. Please report any quality issues you encounter to [email protected], referencing the URI of the item.Includes bibliographical references: p. 76-78.Issued also on microfiche from Lange Micrographics.The objective of this thesis is to examine the feasibility of parallelizing a previously published sequential algorithm. This algorithm uses invariant imbedding to solve an inverse problem associated with the first order, partial differential equation describing radiative transfer. The published algorithm applied invariant imbedding methods to the two stream approximation to a one dimensional radiative transfer equation, resulting in an integrodifferential equation that takes multiple scattering events into account. Two different implementations of the above mentioned serial algorithm viz., Layer Peeling and Characteristic Tracing have been previously described, and the characteristic tracing method has been implemented in parallel. For this thesis, the parallel implementation of the layer peeling algorithm was considered. In this context, layer peeling implies that pertinent quantities are known at some layer, say the (i-I)th, which are used to move deeper, into the ith layer. The parallel implementation was considered because of the anticipated speedup in the solution of the problem under consideration. The algorithm was implemented on an NCUBE 2 machine, with a total of sixty-four processors. Two variations of the parallel algorithm were implemented viz., Block Layer Assignment (BLA), and Rotated Individual Layer Assignment (RILA). The results from the parallel implementation were verified for accuracy by solving the direct problem and using its results as input to the inverse solution. The results obtained from the inverse solution were then compared to the input of the direct solution. Several issues specific to parallel computation were considered in the parallel layer peeling implementation and analysis. These included issues such as task allocation, or the total workload breakdown for assignment to different processors, communication of interim results between processors, and synchronization of the computations from different processors. In order to achieve better performance, the parallel implementation was done by using Reflected Gray Codes for processor assignment. Reflected Gray Code ordering ensures that the address of the processor working on any layer differs in at most a single bit from the processor working on the neighboring layer, so the communication is always between nearest neighbors

Studies on the leaf spot disease mulberry (Morus spp.)

Despite congenial climatic conditions the silk production in the State of Jammu and Kashmir has drastically declined from 76 tonnes in 1980 to 18 tonnes in 1991 (Anonymous, 1992). One of the factors responsible for the decrease in silk production was silkworm and mulberry disease. Almost all the commercial varieties of mulberry (Morus spp.) are prone to various diseases. The diseases not only reduce the leaf yield but impair its nutritional value. Feeding silkworms with diseased leaves adversely of mulberry is one among them. Studies were therefore, conducted to obtain some basic information of systematic nature of the disease so as to know its incidence, intensity symptomatology, causal fungus, perpetuation and management through cultural practices and fungicides. Survey conducted during 1999, revealed that the disease was prevalent in all the five districts of Kashmir Valley with maximum incidence (66.47%) and intensity (50.03%) in district Anantnag. The minimum disease incidence (17.17%) and intensity (7.91%) was observed in district Srinagar. However, an overall average incidence of 41% and intensity 24% of the disease was observed for the Kashmir Valley. The disease has exhibited symptoms very prominently on leaves and can be easily distinguished in five disease development stages. The disease initiates as cicular brown pin head sized spot, which gradually enlarges into dark brown spots surrounded by yellow zone, with whitish centre, producing conidia within acervulli. The mature spots coalesces into large lesions while passing through other stage of development and defoliates. The fungus was isolated and cultured on potato dextrose agar medium at 25 ±10C. its pathogenicity has been proved. The fungus produced hyaline, cylindrical, straight or curved, septate conidia with truncate base, guttulate cells within an acervullus. The average size of the conidia from culture and from fresh spots on leaves was found to be 47.4 x 4.16 and 45.97 x 3.5 µm, respectively. On the basis o symptomatology and morphological characters of the fungus causing leaf spot disease of mulberry, the causal fungus was identified as Phloeospora maculans (Bereng) Allesch an anamorph of Mycosphaerella mori (Fuckel) Wolf The mode of perpetuation of the fungus has been determined. The fungus perpetuates through winter in the form of dormant mycelium in diseased dead overwintered leaves. The conidial formation was observed during 2nd week of April on these leaves. The conidial production was highest, as recorded, (0.58 lac/cm2) on 8th May with 92.8% germination. No perfect state of the fungus was observed under existing conditions. A study on the effect of spacing and training height, revealed that with the increase in spacing and height there was gradual decrease in incidence and intensity of the disease. The maximum incidence (53.68%) and intensity (36.97%) was recorded at 90 x 45cm spacing and minimum incidence (36.21%) and intensity (23.39%) at 180 x 180 cm spacing. Whereas, in case of training the height, maximum incidence (56.46%) and intensity (33.70%) was recorded at height maintained at 10cm (Bush) and minimum incidence (38.835) and intensity (12.73%) at 150cm height (Tall). Screening of 34 mulberry genotypes during the year 1999 and 2000 revealed that none of the variety was immune to disease. Varieties like Obawase, KNG, Brantul and Senmestsu were categorised as resistant to the disease whereas, Botetul, Takawase and Sepentine were found highly susceptible. However, most of the commercial varities such as Goshoerami, Kokuso-20, 21 and 27 Kanva-2, Limoncina etc were found moderately susceptible. In an in vitro study, fungitoxicants both of systemic and protectant nature were significantly effective against conidial germination of Phloeospora maculans. Carbendazim @ 500 ppm inhibited 100 per cent conidial germination. Wheras, 100 per cent inhibition was obtained with triazoles at higher rate of 1000 ppm. Most of the protectants such as mancozeb, captan and zineb inhibited 100% spore germination when used @ 2000 ppm. whereas, similar effect was obtained with copper oxychloride when used @ 4000 ppm. Logistic Regression Model has been applied for the analysis of LD50 for various doses of fungitoxicants used against conidial germination. Under field conditions triazoles gave better results as compared to the carbendazim and protectant fungicides. Hexaconazole @ 500 ppm gave 88.45% disease control, whereas 81.35 and 78.60 per cent disease control was obtained with the application of carbendazim @ 500 ppm and captan @ 2000 ppm. Silk worms hybrid (NB4 D2 x SH6) were tested for the residual effect of fungitoxicants for their mortality, larval weight, cocoon weight and shell weight. The worms were fed in batches on the leaves sprayed 12 days, 8 days 4 days earlier to the feeding. The fungicides triazoles, carbendazim and captan did not show any adverse effect on the silkworms even they were fed leaves sprayed 4 days earlier to the feeding. Whereas, it was found to be safe with feeding of the leaves sprayed with zineb, mancozeb and copper oxychloride 12 days earlier

Computational analysis on conformational dynamics of bone morphogenetic protein-2 (BMP-2)

<p>BMP-2 is widely used for bone regeneration because of its ability to induce osteoblast differentiation and proliferation. The pharmaceutical application of BMP-2 as bone implant makes the studies on stability and conformational dynamics very relevant as proteins are functional only in their native three-dimensional state. Knowing the factors affecting BMP-2 structure becomes essential for designing bone implants activated by BMP-2. In order to explore the influence of temperature and hydration on protein conformation, we have performed the molecular dynamics (MD) simulations at the time scale of 100 ns with two different force fields. We have examined the dynamic behaviour of BMP-2 monomer and dimer in aqueous medium as well as in vacuum at four different temperatures (300, 350, 400 and 450 K). MD simulation of BMP-2 monomer and dimer in water and vacuum environments shows the major contribution of water in structure stabilization. Temperature of the system affects the secondary structure differently in case of monomer and dimer simulation and the dynamics also depends on the environment viz. vacuum and aqueous. Vacuum simulations show very early loss of the major secondary structure content. On the other hand, BMP-2 monomer and dimer in aqueous environment show the unfolding of α-helix with increasing temperature. This unfolded α-helix is converted into β-sheet at 400 K in monomer of BMP-2. Contrary to this, we did not observe β-sheet formation in dimer BMP-2 even at 450 K indicating that monomers are more aggregation prone entity as compared to dimers of BMP-2.</p

Study of Antibacterial Efficacy of Hybrid Chitosan-Silver Nanoparticles for Prevention of Specific Biofilm and Water Purification

Antibacterial efficacy of silver nanoparticles (Ag NPs) deposited alternatively layer by layer (LBL) on chitosan polymer in the form of a thin film over a quartz plate and stainless steel strip has been studied. An eight-bilayer chitosan/silver (Cs/Ag)8 hybrid was prepared having a known concentration of silver. Techniques such as UV-visible spectroscopy, inductively coupled plasma optical emission spectroscopy (ICP-OES), and atomic force microscopy (AFM) were carried out to understand and elucidate the physical nature of the film. Gram-negative bacteria, Escherichia coli (E. coli), were used as a test sample in saline solution for antibacterial studies. The growth inhibition at different intervals of contact time and, more importantly, the antibacterial properties of the hybrid film on repeated cycling in saline solution have been demonstrated. AFM studies are carried out for the first time on the microbe to know the morphological changes affected by the hybrid film. The hybrid films on aging (3 months) are found to be as bioactive as before. Cytotoxicity experiments indicated good biocompatibility. The hybrid can be a promising bioactive material for the prevention of biofilms specific to E. coli and in purification of water for safe drinking

Effect of the Macromolecular Crowding Agents on the Structure and Function of Human Arginase-I, a Therapeutically Important Enzyme

Macromolecular crowding has been known to influence the structure and function of many enzymes through excluded volume effects and/or soft interactions. Here, we employed two synthetic macromolecular crowders, Dextrans and poly(ethylene glycol)s (PEGs) with varying molecular masses, to examine how they affected the structure and function of a therapeutically important enzyme, human arginase-I that catalyzes the conversion of l-arginine to l-ornithine and urea. Except at greater concentrations of Dextran 200, Dextrans were observed to slightly reduce the enzymatic activity, indicating that they exert their influence mainly through the excluded volume effects. Similar outcomes were seen with PEGs, with the exception of PEG 1000, where the activity decreased with increasing PEG concentrations, showing the maximum effect at a 20 g/L concentration. This finding suggests that the enzyme function is reduced by the soft interactions of this macromolecule with the enzyme, supported by the binding measurement. Secondary and local tertiary structures and thermodynamic stability were also affected, suggesting that PEG 1000 has an impact on the protein’s structure. Furthermore, molecular dynamics simulation studies suggest that the catalytic pocket is disturbed, presumably by the unwinding of neighboring helix 9. As a result, the positioning of nearby Glu277 is altered, which prevents His141 and Glu277 from making contact. This hampers the proton transfer from the catalytic His141 to the intermediate species to form ornithine, a crucial step for the substrate hydrolysis reaction by this arginase. Overall, the knowledge gained from this study might be helpful for understanding how different enzymes work in a crowded/cellular environment